Phosphorylation networks regulating JNK activity in diverse genetic backgrounds.
Cellular signaling networks have evolved to enable swift and accurate responses, even in the face of genetic or environmental perturbation. Thus, genetic screens may not identify all the genes that regulate different biological processes. Moreover, although classical screening approaches have succeeded in providing parts lists of the essential components of signaling networks, they typically do not provide much insight into the hierarchical and functional relations that exist among these components. We describe a high-throughput screen in which we used RNA interference to systematically inhibit two genes simultaneously in 17,724 combinations to identify regulators of Drosophila JUN NH(2)-terminal kinase (JNK). Using both genetic and phosphoproteomics data, we then implemented an integrative network algorithm to construct a JNK phosphorylation network, which provides structural and mechanistic insights into the systems architecture of JNK signaling.
Bakal
Chris
C
Department of Genetics, Harvard Medical School, 77 Avenue Louis Pasteur, Boston, MA 02215, USA. cbakal@receptor.med.harvard.edu
Linding
Rune
R
Llense
Flora
F
Heffern
Elleard
E
Martin-Blanco
Enrique
E
Pawson
Tony
T
Perrimon
Norbert
N
D000465:Algorithms; D000818:Animals; D002460:Cell Line; D019295:Computational Biology; D004330:Drosophila / Q000201:enzymology* / Q000235:genetics; D029721:Drosophila Proteins / Q000235:genetics / Q000378:metabolism*; D031541:Fluorescence Resonance Energy Transfer; D017344:Genes, Insect; D048031:JNK Mitogen-Activated Protein Kinases / Q000235:genetics / Q000378:metabolism*; D020935:MAP Kinase Signaling System; D053858:Metabolic Networks and Pathways; D010766:Phosphorylation; D040901:Proteomics; D034622:RNA Interference; D015398:Signal Transduction
D029721:Drosophila Proteins; D048031:JNK Mitogen-Activated Protein Kinases